Novosphingobium aromaticivorans for ccMA Production from Lignin Biomass
Avery C. Vilbert2* (email@example.com), Wayne S. Kontur2, Derek M. Gille2, Daniel R. Noguera1,2, and Timothy J. Donohue1,2
1Great Lakes Bioenergy Research Center; and 2University of Wisconsin–Madison
This project will discuss Novosphingobium aromaticivorans as a bacterial host for production of the commodity chemical cis,cis-muconic acid (ccMA). It will provide further knowledge of the metabolism of aromatics by N. aromaticivorans, as well as a method to produce commodity chemicals from renewable carbon sources that are generated using green microbial engineering techniques.
Millions of tons of the commodity chemical cis,cis-muconic acid (ccMA) is produced annually from finite fossil fuel sources to produce nylon, polyesters, and other materials (Choi et al. 2020). The establishment of a sustainable bioeconomy hinges on the ability to use renewable sources for production of these in-demand chemicals. Lignin is an under-utilized abundant renewable resource that represents a potential carbon source for bio-based production of valuable chemicals. Due to the heterogeneity of lignin, it is challenging to extract commodity chemicals from lignin using current methods (Beckham et al. 2016). This poster discusses the team’s work to funnel lignin streams into a single compound by leveraging the selectivity of enzymes with metabolic engineering techniques. N. aromaticivorans is an ideal candidate for lignin funneling as it is genetically tractable and has the native ability to catabolize lignin derived oligomers and metabolize multiple lignin monomers simultaneously. Previously, the lab was able to convert biomass aromatics into 2-pyrone-4,6-dicarboxylic acid from an engineered N. aromaticivorans strain (Perez et al. 2019). This poster will discuss both the genetic engineering of N. aromaticivorans metabolic pathway for production of ccMA as well as the activity of enzymes along this metabolic pathway. In particular, it will highlight the activity of putative N. aromaticivorans enzymes involved in decarboxylation of protocatechuic acid to catechol and oxidative ring cleavage of catechol to ccMA. This work directly compares the activity of the putative N. aromaticivorans enzymes in vitro and in vivo with other known homologous enzymes to engineer the most efficient ccMA production strain in N. aromaticivorans.
Beckham, G. T., et al. 2016. “Opportunities and Challenges in Biological Lignin Valorization,” Current Opinion in Biotechnology 42, 40–53.
Choi, S., et al. 2020. “Recent Advances in Microbial Production of Cis,cis-Muconic Acid,” Biomolecules 10, 1–14.
Perez, J. M., et al. 2019. “Funneling Aromatic Products of Chemically Depolymerized Lignin into 2-pyrone-4-6-dicarboxylic Acid with Novosphingobium aromaticivorans,” Green Chemistry 21, 1340–50.
This material is based upon work supported by the Great Lakes Bioenergy Research Center, U.S. Department of Energy, Office of Science, Biological and Environmental Research (BER) Program under Award Number DE-SC0018409.